Hepatitis C virus inhibitors

ABSTRACT

The present invention discloses compounds of Formula (I), or pharmaceutically acceptable salts, esters, or prodrugs thereof: 
                         
which inhibit RNA-containing virus, particularly the hepatitis C virus (HCV). Consequently, the compounds of the present invention interfere with the life cycle of the hepatitis C virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HCV infection. The invention also relates to methods of treating an HCV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention. The present invention relates to novel antiviral compounds represented herein above, pharmaceutical compositions comprising such compounds, and methods for the treatment or prophylaxis of viral (particularly HCV) infection in a subject in need of such therapy with the compounds.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application Ser.No. 61/241,617 filed Sep. 11, 2009. The entire teachings of the aboveapplication are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to novel antiviral agents. Morespecifically, the present invention relates to compounds which caninhibit the function of the NS5A protein encoded by Hepatitis C virus(HCV), compositions comprising such compounds, methods for inhibitingHCV viral replication, methods for treating or preventing HCV infection,and processes for making the compounds.

BACKGROUND OF THE INVENTION

Infection with HCV is a major cause of human liver disease throughoutthe world. In the US, an estimated 4.5 million Americans are chronicallyinfected with HCV. Although only 30% of acute infections aresymptomatic, greater than 85% of infected individuals develop chronic,persistent infection. Treatment costs for HCV infection have beenestimated at $5.46 billion for the US in 1997. Worldwide over 200million people are estimated to be infected chronically. HCV infectionis responsible for 40-60% of all chronic liver disease and 30% of allliver transplants. Chronic HCV infection accounts for 30% of allcirrhosis, end-stage liver disease, and liver cancer in the U.S. The CDCestimates that the number of deaths due to HCV will minimally increaseto 38,000/year by the year 2010.

Due to the high degree of variability in the viral surface antigens,existence of multiple viral genotypes, and demonstrated specificity ofimmunity, the development of a successful vaccine in the near future isunlikely. Alpha-interferon (alone or in combination with ribavirin) hasbeen widely used since its approval for treatment of chronic HCVinfection. However, adverse side effects are commonly associated withthis treatment: flu-like symptoms, leukopenia, thrombocytopenia,depression from interferon, as well as anemia induced by ribavirin(Lindsay, K. L. (1997) Hepatology, 26 (suppl 1): 71S-77S). This therapyremains less effective against infections caused by HCV genotype 1(which constitutes ˜75% of all HCV infections in the developed markets)compared to infections caused by the other 5 major HCV genotypes.Unfortunately, only ˜50-80% of the patients respond to this treatment(measured by a reduction in serum HCV RNA levels and normalization ofliver enzymes) and, of responders, 50-70% relapse within 6 months ofcessation of treatment. Recently, with the introduction of pegylatedinterferon (Peg-IFN), both initial and sustained response rates haveimproved substantially, and combination treatment of Peg-IFN withribavirin constitutes the gold standard for therapy. However, the sideeffects associated with combination therapy and the impaired response inpatients with genotype 1 present opportunities for improvement in themanagement of this disease.

First identified by molecular cloning in 1989 (Choo, Q-L et al (1989)Science, 244:359-362), HCV is now widely accepted as the most commoncausative agent of post-transfusion non-A, non-B hepatitis (NANBH) (Kuo,G et al (1989) Science, 244:362-364). Due to its genome structure andsequence homology, this virus was assigned as a new genus in theFlaviviridae family. Like the other members of the Flaviviridae, such asflaviviruses (e.g. yellow fever virus and Dengue virus types 1-4) andpestiviruses (e.g. bovine viral diarrhea virus, border disease virus,and classic swine fever virus) (Choo, Q-L et al (1989) Science,244:359-362; Miller, R. H. and R. H. Purcell (1990) Proc. Natl. Acad.Sci., USA 87:2057-2061), HCV is an enveloped virus containing a singlestrand RNA molecule of positive polarity. The HCV genome isapproximately 9.6 kilobases (kb) with a long, highly conserved,noncapped 5′ nontranslated region (NTR) of approximately 340 bases whichfunctions as an internal ribosome entry site (IRES) (Wang C Y et al ‘AnRNA pseudoknot is an essential structural element of the internalribosome entry site located within the hepatitis C virus 5’ noncodingregion' RNA—A Publication of the RNA Society. 1(5): 526-537, 1995 Jul.).This element is followed by a region which encodes a single long openreading frame (ORF) encoding a polypeptide of ˜3000 amino acidscomprising both the structural and nonstructural viral proteins.

Upon entry into the cytoplasm of the cell, this RNA is directlytranslated into a polypeptide of ˜3000 amino acids comprising both thestructural and nonstructural viral proteins. This large polypeptide issubsequently processed into the individual structural and nonstructuralproteins by a combination of host and virally-encoded proteinases (Rice,C. M. (1996) in B. N. Fields, D. M. Knipe and P. M. Howley (eds)Virology, 2^(nd) Edition, p931-960; Raven Press, N.Y.). There are threestructural proteins, C, E1 and E2. The P7 protein is of unknown functionand is comprised of a highly variable sequence. There are severalnon-structural proteins. NS2 is a zinc-dependent metalloproteinase thatfunctions in conjunction with a portion of the NS3 protein. NS3incorporates two catalytic functions (separate from its association withNS2): a serine protease at the N-terminal end, which requires NS4A as acofactor, and an ATP-ase-dependent helicase function at the carboxylterminus. NS4A is a tightly associated but non-covalent cofactor of theserine protease. NS5A is a membrane-anchored phosphoprotein that isobserved in basally phosphorylated (56 kDa) and hyperphosphorylated (58kDa) forms. While its function has not fully been elucidated, NS5A isbelieved to be important in viral replication. The NS5B protein (591amino acids, 65 kDa) of HCV (Behrens, S. E. et al (1996) EMBO J., 1512-22), encodes an RNA-dependent RNA polymerase (RdRp) activity andcontains canonical motifs present in other RNA viral polymerases. TheNS5B protein is fairly well conserved both intra-typically (˜95-98%amino acid (aa) identity across 1b isolates) and inter-typically (˜85%aa identity between genotype 1a and 1b isolates). The essentiality ofthe HCV NS5B RdRp activity for the generation of infectious progenyvirions has been formally proven in chimpanzees (A. A. Kolykhalov et al.(2000) Journal of Virology, 74(4): 2046-2051). Thus, inhibition of NS5BRdRp activity (inhibition of RNA replication) is predicted to be usefulto treat HCV infection.

Following the termination codon at the end of the long ORF, there is a3′ NTR which roughly consists of three regions: an ˜40 base region whichis poorly conserved among various genotypes, a variable lengthpoly(U)/polypyrimidine tract, and a highly conserved 98 base elementalso called the “3′ X-tail” (Kolykhalov, A. et al (1996) J. Virology,70:3363-3371; Tanaka, T. et al (1995) Biochem Biophys. Res. Commun.,215744-749; Tanaka, T. et al (1996) J. Virology, 70:3307-3312; Yamada,N. et al (1996) Virology, 223:255-261). The 3′ NTR is predicted to forma stable secondary structure which is essential for HCV growth in chimpsand is believed to function in the initiation and regulation of viralRNA replication.

Compounds useful for treating HCV-infected patients are desired whichselectively inhibit HCV viral replication. In particular, compoundswhich are effective to inhibit the function of the NS5A protein aredesired. The HCV NS5A protein is described, for example, in Tan, S.-L.,Katzel, M. G. Virology, 2001, 284, 1; and in Rice, C. M. Nature, 2005,435, 374.

Based on the foregoing, there exists a significant need to identifycompounds with the ability to inhibit HCV.

SUMMARY OF THE INVENTION

The present invention relates to novel antiviral compounds representedherein below, pharmaceutical compositions comprising such compounds, andmethods for the treatment or prophylaxis of viral (particularly HCV)infection in a subject in need of such therapy with said compounds.Compounds of the present invention interfere with the life cycle of thehepatitis C virus and are also useful as antiviral agents.

In its principal aspect, the present invention provides a compound ofFormula (I):

or a pharmaceutically acceptable salt thereof, wherein:

Ring A is absent or a monocyclic or polycyclic group independentlyselected from aryl, heteroaryl, heterocyclic, C₃-C₈ cycloalkyl, andC₃-C₈ cycloalkenyl, each optionally substituted; preferably optionallysubstituted aryl or optionally substituted heteroaryl;

Ring B is a monocyclic or polycyclic group independently selected fromaryl, heteroaryl, heterocyclic, C₃-C₈ cycloalkyl, and C₃-C₈cycloalkenyl, each optionally substituted; preferably optionallysubstituted aryl or optionally substituted heteroaryl;

L is absent or selected from the group consisting of optionallysubstituted C₁-C₄ alkyl, optionally substituted C₂-C₄ alkenyl, andoptionally substituted C₂-C₄ alkynyl;

R¹ at each occurrence is independently hydrogen or optionallysubstituted C₁-C₄ alkyl;

R⁶ at each occurrence is independently selected from the groupconsisting of optionally substituted O(C₁-C₈ alkyl); optionallysubstituted amino; C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₈cycloalkyl, C₃-C₈ cycloalkenyl, heterocyclic, aryl, and heteroaryl, eachoptionally substituted; preferably optionally substituted C₁-C₈ alkyl;more preferably C₁-C₈ alkyl optionally substituted with amino, hydroxy,protected amino or O(C₁-C₄ alkyl);

U is absent or independently selected from O, S, S(O), SO₂, NC(O)—(C₁-C₄alkyl), C(O), protected carbonyl, OCH₂, OCH₂CH₂, SCH₂, SCH₂CH₂, C(R⁷)₂,and C(R⁷)₂C(R⁷)₂; preferably CH₂;

R⁷ at each occurrence is independently selected from the groupconsisting of hydrogen, halogen, cyano, hydroxy, O(C₁-C₄ alkyl), S(C₁-C₄alkyl), amino optionally substituted with one or two C₁-C₄ alkyl,optionally substituted aryl, optionally substituted heteroaryl, andoptionally substituted C₁-C₄ alkyl; preferably hydrogen, halogen orhydroxy;

Optionally two geminal R⁷ groups can be taken together with the carbonatom to which they are attached to form a spiro, optionally substituted3- to 8-membered ring selected from the group consisting of C₃-C₈cycloalkyl, C₃-C₈ cycloalkenyl and 3- to 8-membered heterocyclic, eachoptionally substituted; preferably an optionally substituted C₃-C₈cyclopropyl or an optionally substituted 5- to 6-membered heterocyclic;

R^(7a) and R^(7b) at each occurrence are each independently selectedfrom the group consisting of hydrogen, optionally substituted aryl, andoptionally substituted C₁-C₄ alkyl; alternatively, CHR^(7a)—U orCHR^(7b)—U can be taken together to form a group selected from CH═CH,optionally substituted C₃-C₈ cycloalkyl, optionally substituted aryl,and optionally substituted heterocyclic;

G is optionally substituted 5-membered heteroaryl or optionallysubstituted 5/6-member fused heteroaryl; wherein the 5-memberedheteroaryl contains one or more nitrogen atoms, and wherein the6-membered ring of said 5/6-fused membered heteroaryl is attached to oneof Ring A, L and Ring B, and is aryl or heteroaryl; preferablyoptionally substituted imidazolyl or optionally substitutedbenzimidazolyl;

Q is selected from:

Alternatively G and Q are taken together to form

V is selected from the group consisting of —N(R^(1c))—,—N(R^(1c))—C(O)—, —N(R^(1c))—C(O)O—, and —N(R^(1c))—C(O)—N(R^(1c))—;preferably —N(R^(1c))— or —N(R^(1c))—C(O)—;

W is O or —N(R^(1b))—; preferably —N(R^(1b))—;

R^(1a), R^(1b), R^(1c), R^(1d), R⁹, and R^(9a) are each independentlyhydrogen or optionally substituted C₁-C₄ alkyl; alternatively R^(1a) andR^(9a), R^(1a) and R⁹, R^(1d) and R^(9a), or R^(1d) and R⁹ can be takentogether with the carbon atom(s) to which they are attached to form anoptionally substituted C₃-C₈ cycloalkyl or optionally substituted 4- to8-membered heterocyclic; or yet alternatively R^(1b) and R^(9a), orR^(1b) and R⁹ can be taken together with the nitrogen or carbon atom(s)to which they are attached to form an optionally substituted 4- to8-membered heterocyclic;

X is absent, O, S, CH₂, or CH₂CH₂;

Y is absent, O, S, C(R¹)₂, C(R¹)₂C(R⁷)₂, C(R¹)₂C(R⁷)₂C(R⁷)₂, C(R¹)₂,C(R¹)₂, or C(R¹)₂SC(R¹)₂;

Wherein at least one of X and Y is present;

Wherein at least one of X and Y is not O or S;

R² is hydrogen, optionally substituted C₁-C₈ alkyl, or —NR^(a)R^(b);

R^(a) is hydrogen or optionally substituted C₁-C₈ alkyl;

R^(b) at each occurrence is —C(O)—R⁶;

Alternatively R^(a) and R^(b) can be taken together with the nitrogenatom to which they are attached to form an optionally substitutedheterocyclic or optionally substituted heteroaryl group;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₈ alkyl, optionally substitutedC₂-C₈ alkenyl, and optionally substituted C₃-C₈ cycloalkyl; preferablyhydrogen or optionally substituted C₁-C₄ alkyl; alternatively, R³ and R⁴can be taken together with the carbon atom to which they are attached toform optionally substituted C₃-C₈ cycloalkyl or optionally substitutedheterocyclic; and

R⁵ is independently hydrogen, optionally substituted C₁-C₈ alkyl, oroptionally substituted C₃-C₈ cycloalkyl; preferably hydrogen oroptionally substituted C₁-C₄ alkyl.

Each preferred group stated above can be taken in combination with one,any or all other preferred groups.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundor combination of compounds of the present invention, or apharmaceutically acceptable salt thereof, in combination with apharmaceutically acceptable carrier or excipient.

In yet another aspect, the present invention provides a method ofinhibiting the replication of a RNA-containing virus comprisingcontacting said virus with said pharmaceutical composition.Particularly, this invention is directed to methods of inhibiting thereplication of HCV.

In still another aspect, the present invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising administering to a patient in need of such treatment atherapeutically effective amount of a compound or combination ofcompounds of the present invention, or a pharmaceutically acceptablesalt thereof. Particularly, this invention is directed to methods oftreating or preventing infection caused by HCV.

Yet another aspect of the present invention provides the use of acompound or combination of compounds of the present invention, or apharmaceutically acceptable salt thereof, as defined hereinafter, in thepreparation of a medicament for the treatment or prevention of infectioncaused by RNA-containing virus, specifically HCV.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds of Formula (I) as illustratedabove, or a pharmaceutically acceptable salt thereof.

The compounds of the invention have utility in inhibiting thereplication of RNA-containing virus, including, for example, HCV. Othercompounds useful for inhibiting the replication of RNA-containingviruses and/or for the treatment or prophylaxis of HCV infection havebeen described in copending U.S. application Ser. No. 12/702,673 filedFeb. 9, 2010 entitled “Linked Dibenzimidiazole Derivatives”; U.S.application Ser. No. 12/702,692 filed Feb. 9, 2010 entitled “LinkedDibenzimidiazole Derivatives”; U.S. application Ser. No. 12/702,802filed Feb. 9, 2010 entitled “Linked Dibenzimidiazole Derivatives”; U.S.application Ser. No. 12/707,190 filed Feb. 17, 2010 entitled “LinkedDiimidazole Antivirals”; U.S. application Ser. No. 12/707,200 filed Feb.17, 2010 entitled “Linked Diimidazole Derivatives”; U.S. applicationSer. No. 12/707,210 filed Feb. 17, 2010 entitled “Hepatitis C VirusInhibitors”; U.S. application Ser. No. 12/714,583 filed Mar. 1, 2010entitled “Novel Benzimidazole Derivatives”; and U.S. application Ser.No. 12/714,576 filed Mar. 1, 2010 entitled “Hepatitis C VirusInhibitors”; U.S. application Ser. No. 12/816,148 filed Jun. 15, 2010entitled “Hepatitis C Virus Inhibitors”; U.S. application Ser. No.12/816,171 filed Jun. 15, 2010 entitled “Hepatitis C Virus Inhibitors”;U.S. Provisional Application Ser. No. 61/241,489 filed Sep. 11, 2009entitled “Hepatitis C Virus Inhibitors”; U.S. Provisional ApplicationSer. No. 61/241,578 filed Sep. 11, 2009 entitled “Hepatitis C VirusInhibitors”; U.S. Provisional Application Ser. No. 61/241,595 filed Sep.11, 2009 entitled “Hepatitis C Virus Inhibitors”; U.S. ProvisionalApplication Ser. No. 61/241,617 filed Sep. 11, 2009 entitled “HepatitisC Virus Inhibitors”; U.S. Provisional Application Ser. No. 61/241,577filed Sep. 11, 2009 entitled “Hepatitis C Virus Inhibitors”; U.S.Provisional Application Ser. No. 61/241,598 filed Sep. 11, 2009 entitled“Hepatitis C Virus Inhibitors”; U.S. Provisional Application Ser. No.61/286,178 filed Dec. 14, 2009 entitled “Hepatitis C Virus Inhibitors”;U.S. Provisional Application Ser. No. 61/297,918 filed Jan. 25, 2010entitled “Hepatitis C Virus Inhibitors”; U.S. Provisional ApplicationSer. No. 61/314,304 filed Mar. 16, 2010 entitled “Hepatitis C VirusInhibitors”; U.S. Provisional Application Ser. No. 61/322,438 filed Apr.9, 2010 entitled “Hepatitis C Virus Inhibitors”; U.S. ProvisionalApplication Ser. No. 61/351,327 filed Jun. 4, 2010 entitled “Hepatitis CVirus Inhibitors”; U.S. Provisional Application Ser. No. 61/372,999filed Aug. 12, 2010 entitled “Hepatitis C Virus Inhibitors”; and thecontents of each of which are expressly incorporated by referenceherein.

As discussed above, a general strategy for the development of antiviralagents is to inactivate virally encoded proteins, including NS5A, thatare essential for the replication of the virus. The relevant patentdisclosures describing the synthesis of HCV NS5A inhibitors are: US2009/0202478; US 2009/0202483; WO 2004/014852; WO 2006/079833; WO2006/1333262; WO 2007/031791; WO 2007/070556; WO 2007/070600; WO2007/082554; WO 2008/021927; WO 2008/021928; WO 2008/021936; WO2008/048589; WO 2008/064218; WO 2008/070447; WO 2008/144380; WO2008/154601; WO 2009/020825; WO 2009/020828; WO 2009/034390; WO2009/102318; WO 2009/102325; WO 2009/102694; WO 2010/017401; WO2010/039793; WO 2010/065668; WO 2010/065674; WO 2010/065681; WO2010/091413; WO 2010/096777; WO 2010/096462 and WO 2010/096302, thecontents of each of which are expressly incorporated by referenceherein.

In still another embodiment, the present invention relates to compoundsof Formula (I), and pharmaceutically acceptable salts thereof; whereintwo geminal R⁷ groups, taken together with the carbon atom to which theyare attached, form a spiro, optionally substituted 3- to 8-membered ringselected from the group consisting of C₃-C₈ cycloalkyl, C₃-C₈cycloalkenyl and 3- to 8-membered heterocyclic, each optionallysubstituted.

In still another embodiment, the present invention relates to compoundsof Formula (I), and pharmaceutically acceptable salts thereof; whereintwo geminal R⁷ groups, taken together with the carbon atom to which theyare attached, form a spiro, optionally substituted cyclopropyl or aspiro, optionally substituted 5- to 6-membered heterocyclic.

In another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein

is one of the following groups:

each of which can be optionally substituted.

In yet another embodiment, R^(1a) and R^(9a), R^(1a) and R⁹, R^(1d) andR^(9a), or R^(1d) and R⁹ are taken together with the carbon atoms towhich they are attached to form an optionally substituted C₃-C₈cycloalkyl or optionally substituted 4- to 8-membered heterocyclic.

In a further embodiment R^(1b) and R^(9a), or R^(1b) and R⁹ are takentogether with the nitrogen or carbon atom(s) to which they are attachedto form an optionally substituted 4- to 8-membered heterocyclic.

In yet another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein themoiety of

is one of the following core groups:

wherein R⁶ is as previously defined and each of the above core groups isoptionally substituted.

In an additional embodiment, the invention relates to compounds ofFormula (I) and pharmaceutically acceptable salts thereof; wherein R₆ isindependently an optionally substituted C₁-C₈ alkyl.

In a further embodiment, the invention relates to compounds of Formula(I) and pharmaceutically acceptable salts thereof; wherein —C(O)R₆ is anα-amino acid residue.

In yet another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein G isan optionally substituted five-membered heteroaryl containing one ormore nitrogen atoms, and is C-attached.

In yet another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein G isan optionally substituted 5/6-membered fused heteroaryl; wherein the5-membered ring of said 5/6-membered fused heteroaryl is a heteroarylcontaining one or more nitrogen atoms and wherein the 5-membered ring isC-attached to group Q, and wherein the 6-membered ring of said5/6-membered fused heteroaryl is aryl or heteroaryl and is C-attached toone of Ring A, L and Ring B.

In yet another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein G isone of the following heteroaryl groups:

wherein each of the above heteroaryl groups is optionally substituted.

In yet another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof; wherein G isoptionally substituted imidazolyl or benzimidazolyl.

In yet another embodiment, the present invention relates to compounds ofFormulae (Ia-1˜Ia-4), and pharmaceutically acceptable salts thereof:

wherein Ring A, Ring B, G, J, L, U, W, X, Y, R¹, R², R³, R⁴, R⁵, R⁶, R⁹,R^(1a), R^(1b), R^(1d), R^(7a), R^(7b) and R^(9a) are as previouslydefined.

In yet another embodiment, the present invention relates to compounds ofFormulae (Ia-5˜Ia-8), and pharmaceutically acceptable salts thereof:

wherein Ring A, Ring B, G, J, L, U, W, X, Y, R¹, R², R³, R⁴, R⁵, R⁶, R⁹,R^(1a), R^(2b), R^(1c), R^(1d), R^(7a), R^(7b) and R^(9a) are aspreviously defined.

In yet another embodiment, the present invention relates to compounds ofFormulae (Ia-9˜Ia-12), and pharmaceutically acceptable salts thereof:

wherein Ring A, Ring B, G, L, R⁶, R^(7a) and R^(7b) are as previouslydefined.

In yet another embodiment, the present invention relates to compounds ofFormulae (Ia-13˜Ia-16), and pharmaceutically acceptable salts thereof:

wherein Ring A, Ring B, G, L, R⁶, R^(7a) and R^(7b) are as previouslydefined.

In yet another embodiment, the present invention relates to compounds ofFormulae (Ia-13˜Ia-16), and pharmaceutically acceptable salts thereof;wherein R⁶ at each occurrence is selected from the group consisting ofC₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₈ cycloalkyl, C₃-C₈cycloalkenyl, heterocyclic, aryl, and heteroaryl, each optionallysubstituted.

In still another embodiment, the present invention relates to compoundsof Formula (Ia-13˜Ia-16), and pharmaceutically acceptable salts thereof;wherein R⁶ at each occurrence is independently C₁-C₈ alkyl optionallysubstituted with amino, hydroxy, protected amino, or O(C₁-C₄ alkyl).

In still another embodiment, the present invention relates to compoundsof Formulae (Ib-1˜Ib-4), and pharmaceutically acceptable salts thereof:

wherein Ring B, G, Q, U, R¹, R⁶, R^(7a) and R^(7b) are as previouslydefined; in Formula (Ib-1), Ring A and L are each present and aspreviously defined; in Formula (Ib-2), Ring A is present and aspreviously defined; and in Formula (Ib-3), L is present and aspreviously defined.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-1˜Ib-4), and pharmaceutically acceptable salts thereof;wherein R⁶ is independently C₁-C₈ alkyl optionally substituted withamino, hydroxy, protected amino or O(C₁-C₄ alkyl).

In still another embodiment, the present invention relates to compoundsof Formula (Ib-1), and pharmaceutically acceptable salts thereof;wherein Ring A and Ring B are each independently optionally substitutedphenyl or monocyclic heteroaryl; L is optionally substituted C₂-C₄alkenyl or optionally substituted C₂-C₄ alkynyl; and G is optionallysubstituted imidazolyl or benzimidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-2), and pharmaceutically acceptable salts thereof;wherein one of Ring A and Ring B is optionally substituted phenyl oroptionally substituted monocyclic heteroaryl, the other of Ring A andRing B is optionally substituted bicyclic aryl or heteroaryl; and G isoptionally substituted imidazolyl or benzimidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-2), and pharmaceutically acceptable salts thereof;wherein Ring A and Ring B are each independently optionally substitutedbicyclic aryl or heteroaryl; and G is optionally substituted imidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-2), and pharmaceutically acceptable salts thereof;wherein Ring A and Ring B are each independently optionally substitutedphenyl or monocyclic heteroaryl; and G is optionally substitutedbenzimidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-3), and pharmaceutically acceptable salts thereof;wherein Ring B is optionally substituted bicyclic aryl or heteroaryl; Lis optionally substituted C₂-C₄ alkenyl or optionally substituted C₂-C₄alkynyl; and G is optionally substituted imidazolyl or benzimidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-3), and pharmaceutically acceptable salts thereof;wherein B is optionally substituted phenyl or monocyclic heteroaryl; Lis optionally substituted C₂-C₄ alkenyl or optionally substituted C₂-C₄alkynyl; and G is optionally substituted imidazolyl or benzimidazolyl.

In still another embodiment, the present invention relates to compoundsof Formula (Ib-4), and pharmaceutically acceptable salts thereof;wherein B is optionally substituted polycyclic aryl or heteroaryl; and Gis optionally substituted imidazolyl or benzimidazolyl.

Representative compounds of the present invention are those selectedfrom compounds I-348 compiled in the following tables:

TABLE 1 Compounds 1-219.

Entry

 1

 2

 3

 4

 5

 6

 7

 8

 9

 10

 11

 12

 13

 14

 15

 16

 17

 18

 19

 20

 21

 22

 23

 24

 25

 26

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 28

 29

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 31

 32

 33

 34

 35

 36

 37

 38

 39

 40

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 42

 43

 44

 45

 46

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 48

 49

 50

 51

 52

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 54

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 58

 59

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 61

 62

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 64

 65

 66

 67

 68

 69

 70

 71

 72

 73

 74

 75

 76

 77

 78

 79

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 81

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 95

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 97

 98

 99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215

216

217

218

219

TABLE 2 Compounds 220-229.

Entry R R′ R″ U Entry R R′ R″ U 220 Me H H CH₂ 221 H H H CF₂ 222 Me H HS 223 H H H

224 Me H H O 225 H H H

226 H Ph H CH₂ 227 H H H

228 H H Ph CH₂ 229 H H H

TABLE 3 Compounds 230-233.

Entry E Entry E Entry E Entry E 230 absent 231 CH₂ 232 CH₂O 233 OCH₂CH₂

TABLE 4 Compounds 234-259.

Entry R 234

235

236

237

238

239

240

241

242

243

244

245

246

247

248

249

250

251

252

253

254

255

256

257

258

259

TABLE 5 Compounds 260-268.

Entry Q^(q) 260

261

262

263

264

265

266

267

268

TABLE 6 Compounds 269-278.

Entry R R′ R″ R′′′ Entry R R′ R″ R′′′ 269 F H H H 270 F F H H 271 Me H HH 272 Me Me H H 273 H H Me Me 274 H H Et Et 275 CF₃ H H H 276 CF₃ H CF₃H 277 Cl H H H 278 Cl H Cl H

TABLE 7 Compounds 279-296.

Entry R R′ R″ Entry R R′ R″ 279 Me H H 280 H CO₂H H 281 H F H 282 H HCO₂H 283 H H F 284 H CO₂Me H 285 H Cl H 286 H H CO₂Me 287 H H Cl 288 HCONH₂ H 289 H Me H 290 H H CONH₂ 291 H H Me 292 H OMe H 293 H CF₃ H 294H H OMe 295 H H CF₃ 296 CO₂Me H H

Table8: Compounds 297-314.

Entry A^(a) 297

298

299

300

301

302

303

304

305

306

307

308

309

310

311

312

313

314

TABLE 9 Compounds 315-335.

Entry A^(a) 315

316

317

318

319

320

321

322

323

324

325

326

327

328

329

330

331

332

333

334

335

TABLE 10 Compounds 336-347.

Entry G^(g) Entry G^(g) 336

337

338

339

340

341

342

343

344

345

346

347

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principals ofchemical bonding. In some instances it may be necessary to remove ahydrogen atom in order to accommodate a substitutent at any givenlocation.

It is intended that the definition of any substituent or variable (e.g.,R³, R⁷, etc.) at a particular location in a molecule be independent ofits definitions elsewhere in that molecule. For example, when U isC(R⁷)₂, each of the two R⁷ groups may be the same or different.

It is also intended that the drawn formula of any linker or spacer(e.g., —NH—C(O)—, —OC(O)—NH—, etc.) at a particular location in amolecule be independent of its directions of attachment, unlessotherwise indicated. For example, when a linker is —NH—C(O)—, it ismeant to include —NH—C(O)— and —C(O)—NH— for attachments.

It is also intended that in some embodiments, the compounds of thepresent invention may contain one or more asymmetric carbon atoms andmay exist in racemic, diastereoisomeric, and optically active forms. Itwill still be appreciated that certain compounds of the presentinvention may exist in different tautomeric forms. All tautomers arecontemplated to be within the scope of the present invention.

It should be understood that the compounds encompassed by the presentinvention are those that are suitably stable for use as pharmaceuticalagent.

It will be further appreciated that reference herein to therapy and/ortreatment includes, but is not limited to, prevention, retardation,prophylaxis, therapy and/or cure of the disease. It will further beappreciated that references herein to treatment or prophylaxis of HCVinfection includes treatment or prophylaxis of HCV-associated diseasesuch as liver fibrosis, cirrhosis and hepatocellular carcinoma.

A further embodiment of the present invention includes pharmaceuticalcompositions comprising any single compound a combination of two or morecompounds delineated herein, and pharmaceutically acceptable saltsthereof, with a pharmaceutically acceptable carrier or excipient.

Yet a further embodiment of the present invention is a pharmaceuticalcomposition comprising any single compound or a combination of two ormore compounds delineated herein, and pharmaceutically acceptable saltsthereof, in combination with one or more agents known in the art, with apharmaceutically acceptable carrier or excipient.

It will be further appreciated that compounds of the present inventioncan be administered as the sole active pharmaceutical agent, or used incombination with one or more agents to treat or prevent hepatitis Cinfections or the symptoms associated with HCV infection. Other agentsto be administered in combination with a compound or combination ofcompounds of the present invention include therapies for disease causedby HCV infection that suppresses HCV viral replication by direct orindirect mechanisms. These agents include, but not limited to, hostimmune modulators (for example, interferon-alpha, pegylatedinterferon-alpha, consensus interferon, interferon-beta,interferon-gamma, CpG oligonucleotides and the like); antiviralcompounds that inhibit host cellular functions such as inosinemonophosphate dehydrogenase (for example, ribavirin and the like);cytokines that modulate immune function (for example, interleukin 2,interleukin 6, and interleukin 12); a compound that enhances thedevelopment of type 1 helper T cell response; interfering RNA;anti-sense RNA; vaccines comprising HCV antigens or antigen adjuvantcombinations directed against HCV; agents that interact with hostcellular components to block viral protein synthesis by inhibiting theinternal ribosome entry site (IRES) initiated translation step of HCVviral replication or to block viral particle maturation and release withagents targeted toward the viroporin family of membrane proteins suchas, for example, HCV P7 and the like; and any agent or combination ofagents that inhibit the replication of HCV by targeting other proteinsof the viral genome involved in the viral replication and/or interferewith the function of other viral targets, such as inhibitors of NS3/NS4Aprotease, NS3 helicase, NS5B polymerase, NS4A protein and NS5A protein.

According to yet another embodiment, the pharmaceutical compositions ofthe present invention may further comprise other inhibitor(s) of targetsin the HCV life cycle, including, but not limited to, helicase,polymerase, metalloprotease, NS4A protein, NS5A protein, and internalribosome entry site (IRES).

Accordingly, one embodiment of the present invention is directed to amethod for treating or preventing an infection caused by anRNA-containing virus comprising co-administering to a patient in need ofsuch treatment one or more agents selected from the group consisting ofa host immune modulator and a second or more antiviral agents, or acombination thereof, with a therapeutically effective amount of acompound or combination of compounds of the present invention, or apharmaceutically acceptable salt thereof. Examples of the host immunemodulator include, but are not limited to, interferon-alpha,pegylated-interferon-alpha, interferon-beta, interferon-gamma, acytokine, a vaccine, and a vaccine comprising an antigen and anadjuvant, and said second antiviral agent inhibits replication of HCVeither by inhibiting host cellular functions associated with viralreplication or by targeting proteins of the viral genome. An example ofthe RNA-containing virus includes, but is not limited to, hepatitis Cvirus (HCV).

A further embodiment of the present invention is directed to a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment anagent or combination of agents that treat or alleviate symptoms of HCVinfection including cirrhosis and inflammation of the liver, with atherapeutically effective amount of a compound or combination ofcompounds of the present invention, or a pharmaceutically acceptablesalt thereof. An example of the RNA-containing virus includes, but isnot limited to, hepatitis C virus (HCV).

Yet another embodiment of the present invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents that treat patients for disease caused by hepatitis B(HBV) infection, with a therapeutically effective amount of a compoundor a combination of compounds of the present invention, or apharmaceutically acceptable salt thereof. An agent that treats patientsfor disease caused by hepatitis B (HBV) infection may be for example,but not limited thereto, L-deoxythymidine, adefovir, lamivudine ortenfovir, or any combination thereof. An example of the RNA-containingvirus includes, but is not limited to, hepatitis C virus (HCV).

A further embodiment of the present invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents that treat patients for disease caused by humanimmunodeficiency virus (HIV) infection, with a therapeutically effectiveamount of a compound or a combination of compounds of the presentinvention, or a pharmaceutically acceptable salt thereof. The agent thattreats patients for disease caused by human immunodeficiency virus (HIV)infection may include, but is not limited thereto, ritonavir, lopinavir,indinavir, nelfinavir, saquinavir, amprenavir, atazanavir, tipranavir,TMC-114, fosamprenavir, zidovudine, lamivudine, didanosine, stavudine,tenofovir, zalcitabine, abacavir, efavirenz, nevirapine, delavirdine,TMC-125, L-870812, S-1360, enfuvirtide (T-20) or T-1249, or anycombination thereof. An example of the RNA-containing virus includes,but is not limited to, hepatitis C virus (HCV).

It can occur that a patient may be co-infected with hepatitis C virusand one or more other viruses, including, but not limited to, humanimmunodeficiency virus (HIV), hepatitis A virus (HAV) and hepatitis Bvirus (HBV). Thus also contemplated is combination therapy to treat suchco-infections by co-administering a compound according to the presentinvention with at least one of an HIV inhibitor, an HAV inhibitor and anHBV inhibitor.

In addition, the present invention provides the use of a compound or acombination of compounds of the invention, or a pharmaceuticallyacceptable salt thereof, and one or more agents selected from the groupconsisting of a host immune modulator and a second or more antiviralagents, or a combination thereof, to prepare a medicament for thetreatment of an infection caused by an RNA-containing virus in apatient, particularly hepatitis C virus. Examples of the host immunemodulator are, but not limited to, interferon-alpha,pegylated-interferon-alpha, interferon-beta, interferon-gamma, acytokine, a vaccine, and a vaccine comprising an antigen and anadjuvant, and said second antiviral agent inhibits replication of HCVeither by inhibiting host cellular functions associated with viralreplication or by targeting proteins of the viral genome.

When used in the above or other treatments, combination of compound orcompounds of the present invention, together with one or more agents asdefined herein above, can be employed in pure form or, where such formsexist, in pharmaceutically acceptable salt thereof. Alternatively, suchcombination of therapeutic agents can be administered as apharmaceutical composition containing a therapeutically effective amountof the compound or combination of compounds of interest, or theirpharmaceutically acceptable salt thereof, in combination with one ormore agents as defined hereinabove, and a pharmaceutically acceptablecarrier. Such pharmaceutical compositions can be used for inhibiting thereplication of an RNA-containing virus, particularly Hepatitis C virus(HCV), by contacting said virus with said pharmaceutical composition. Inaddition, such compositions are useful for the treatment or preventionof an infection caused by an RNA-containing virus, particularlyHepatitis C virus (HCV).

Hence, a still further embodiment of the invention is directed to amethod of treating or preventing infection caused by an RNA-containingvirus, particularly a hepatitis C virus (HCV), comprising administeringto a patient in need of such treatment a pharmaceutical compositioncomprising a compound or combination of compounds of the invention or apharmaceutically acceptable salt thereof, and one or more agents asdefined hereinabove, with a pharmaceutically acceptable carrier.

When administered as a combination, the therapeutic agents can beformulated as separate compositions which are given at the same time orwithin a predetermined period of time, or the therapeutic agents can begiven as a single unit dosage form.

Antiviral agents contemplated for use in such combination therapyinclude agents (compounds or biologicals) that are effective to inhibitthe formation and/or replication of a virus in a mammal, including butnot limited to agents that interfere with either host or viralmechanisms necessary for the formation and/or replication of a virus ina mammal. Such agents can be selected from another anti-HCV agent; anHIV inhibitor; an HAV inhibitor; and an HBV inhibitor.

Other agents to be administered in combination with a compound of thepresent invention include a cytochrome P450 monooxygenase inhibitor(also referred to herein as a CYP inhibitor), which is expected toinhibit metabolism of the compounds of the invention. Therefore, thecytochrome P450 monooxygenase inhibitor would be in an amount effectiveto inhibit metabolism of the compounds of this invention. Accordingly,the CYP inhibitor is administered in an amount such that thebioavailiablity of the protease inhibitor is increased in comparison tothe bioavailability in the absence of the CYP inhibitor.

In one embodiment, the invention provides methods for improving thepharmacokinetics of compounds of the invention. The advantages ofimproving the pharmacokinetics of drugs are recognized in the art (see,for example, US Patent Pub. Nos. 2004/0091527; US 2004/0152625; and US2004/0091527). Accordingly, one embodiment of this invention provides amethod for administering an inhibitor of CYP3A4 and a compound of theinvention. Another embodiment of this invention provides a method foradministering a compound of the invention and an inhibitor of isozyme3A4 (“CYP3A4”), isozyme 2C19 (“CYP2C19”), isozyme 2D6 (“CYP2D6”),isozyme 1A2 (“CYP1A2”), isozyme 2C9 (“CYP2C9”), or isozyme 2E1(“CYP2E1”). In a preferred embodiment, the CYP inhibitor preferablyinhibits CYP3A4. Any CYP inhibitor that improves the pharmacokinetics ofthe relevant NS3/4A protease may be used in a method of this invention.These CYP inhibitors include, but are not limited to, ritonavir (see,for example, WO 94/14436), ketoconazole, troleandomycin,4-methylpyrazole, cyclosporin, clomethiazole, cimetidine, itraconazole,fluconazole, miconazole, fluvoxamine, fluoxetine, nefazodone,sertraline, indinavir, nelfinavir, amprenavir, fosamprenavir,saquinavir, lopinavir, delavirdine, erythromycin, VX-944, and VX-497.Preferred CYP inhibitors include ritonavir, ketoconazole,troleandomycin, 4-methylpyrazole, cyclosporin, and clomethiazole.

It will be understood that the administration of the combination of theinvention by means of a single patient pack, or patient packs of eachformulation, containing within a package insert instructing the patientto the correct use of the invention is a desirable additional feature ofthis invention.

According to a further aspect of the invention is a pack comprising atleast a compound of the invention and a CYP inhibitor of the inventionand an information insert containing directions on the use of thecombination of the invention. In an alternative embodiment of thisinvention, the pharmaceutical pack further comprises one or more ofadditional agent as described herein. The additional agent or agents maybe provided in the same pack or in separate packs.

Another aspect of this involves a packaged kit for a patient to use inthe treatment of HCV infection or in the prevention of HCV infection,comprising: a single or a plurality of pharmaceutical formulation ofeach pharmaceutical component; a container housing the pharmaceuticalformulation(s) during storage and prior to administration; andinstructions for carrying out drug administration in a manner effectiveto treat or prevent HCV infection.

Accordingly, this invention provides kits for the simultaneous orsequential administration of a compound of the invention and a CYPinhibitor (and optionally an additional agent) or derivatives thereofare prepared in a conventional manner. Typically, such a kit willcomprise, e.g. a composition of each inhibitor and optionally theadditional agent(s) in a pharmaceutically acceptable carrier (and in oneor in a plurality of pharmaceutical formulations) and writteninstructions for the simultaneous or sequential administration.

In another embodiment, a packaged kit is provided that contains one ormore dosage forms for self administration; a container means, preferablysealed, for housing the dosage forms during storage and prior to use;and instructions for a patient to carry out drug administration. Theinstructions will typically be written instructions on a package insert,a label, and/or on other components of the kit, and the dosage form orforms are as described herein. Each dosage form may be individuallyhoused, as in a sheet of a metal foil-plastic laminate with each dosageform isolated from the others in individual cells or bubbles, or thedosage forms may be housed in a single container, as in a plasticbottle. The present kits will also typically include means for packagingthe individual kit components, i.e., the dosage forms, the containermeans, and the written instructions for use. Such packaging means maytake the form of a cardboard or paper box, a plastic or foil pouch, etc.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “viral infection” refers to the introduction of a virus intocells or tissues, e.g., hepatitis C virus (HCV). In general, theintroduction of a virus is also associated with replication. Viralinfection may be determined by measuring virus antibody titer in samplesof a biological fluid, such as blood, using, e.g., enzyme immunoassay.Other suitable diagnostic methods include molecular based techniques,such as RT-PCR, direct hybrid capture assay, nucleic acid sequence basedamplification, and the like. A virus may infect an organ, e.g., liver,and cause disease, e.g., hepatitis, cirrhosis, chronic liver disease andhepatocellular carcinoma.

The term “immune modulator” refers to any substance meant to alter theworking of the humoral or cellular immune system of a subject. Suchimmune modulators include inhibitors of mast cell-mediated inflammation,interferons, interleukins, prostaglandins, steroids, cortico-steroids,colony-stimulating factors, chemotactic factors, etc.

The term “aryl,” as used herein, refers to a mono- or polycycliccarbocyclic ring system including, but not limited to, phenyl, naphthyl,tetrahydronaphthyl, indanyl, and idenyl. A polycyclic aryl is apolycyclic ring system that comprises at least one aromatic ring.Polycyclic aryls can comprise fused rings, covalently attached rings ora combination thereof.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclicring system comprising at least one aromatic ring having one or morering atom selected from S, O and N; and the remaining ring atoms arecarbon, wherein any N or S contained within the ring may be optionallyoxidized. Heteroaryl includes, but is not limited to, pyridinyl,pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl,oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, andquinoxalinyl. A polycyclic heteroaryl can comprise fused rings,covalently attached rings or a combination thereof.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

The term “bicyclic aryl” or “bicyclic heteroaryl” refers to a ringsystem consisting of two rings wherein at least one ring is aromatic;and they can be fused or covalently attached.

The term “tricyclic aryl” or “tricyclic heteroaryl” refers to a ringsystem consisting of three rings wherein at least one ring is aromatic.

The terms “C₁-C₄ alkyl,” “C₁-C₆ alkyl,” “C₁-C₈ alkyl,” “C₂-C₄ alkyl,” or“C₃-C₆ alkyl,” as used herein, refer to saturated, straight- orbranched-chain hydrocarbon radicals containing between one and four, oneand six, one and eight carbon atoms, or the like, respectively. Examplesof C₁-C₈ alkyl radicals include, but are not limited to, methyl, ethyl,propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl andoctyl radicals.

The terms “C₂-C₈ alkenyl,” “C₂-C₄ alkenyl,” “C₃-C₄ alkenyl,” or “C₃-C₆alkenyl,” as used herein, refer to straight- or branched-chainhydrocarbon radicals containing from two to eight, or two to four carbonatoms, or the like, having at least one carbon-carbon double bond by theremoval of a single hydrogen atom. Alkenyl groups include, but are notlimited to, for example, ethenyl, propenyl, butenyl,1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

The terms “C₂-C₈ alkynyl,” “C₂-C₄ alkynyl,” “C₃-C₄ alkynyl,” or “C₃-C₆alkynyl,” as used herein, refer to straight- or branched-chainhydrocarbon radicals containing from two to eight, or two to four carbonatoms, or the like, having at least one carbon-carbon triple bond by theremoval of a single hydrogen atom. Representative alkynyl groupsinclude, but are not limited to, for example, ethynyl, 1-propynyl,1-butynyl, heptynyl, octynyl, and the like.

The term “C₃-C₈-cycloalkyl”, or “C₅-C₇-cycloalkyl,” as used herein,refers to a monocyclic or polycyclic saturated carbocyclic ringcompound, and the carbon atoms may be optionally oxo-substituted.Examples of C₃-C₈-cycloalkyl include, but not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl and cyclooctyl; andexamples of C₅-C₇-cycloalkyl include, but not limited to, cyclopentyl,cyclohexyl, bicyclo[2.2.1]heptyl, and the like.

The term “C₃-C₈ cycloalkenyl” or “C₅-C₇ cycloalkenyl” as used herein,refers to monocyclic or polycyclic carbocyclic ring compound having atleast one carbon-carbon double bond, and the carbon atoms may beoptionally oxo-substituted. Examples of C₃-C₈ cycloalkenyl include, butnot limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like; and examples ofC₅-C₇ cycloalkenyl include, but not limited to, cyclopentenyl,cyclohexenyl, cycloheptenyl, and the like.

The term “arylalkyl”, as used herein, refers to an aryl-substitutedalkyl group. More preferred arylalkyl groups are aryl-C₁-C₆-alkylgroups.

The term “heteroarylalkyl”, as used herein, refers to aheteroaryl-substituted alkyl group. More preferred heteroarylalkylgroups are heteroaryl-C₁-C₆-alkyl groups.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl andcycloalkenyl moiety described herein can also be an aliphatic group oran alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of anycombination of carbon atoms, hydrogen atoms, halogen atoms, oxygen,nitrogen or other atoms, and optionally contains one or more units ofunsaturation, e.g., double and/or triple bonds. Examples of aliphaticgroups are functional groups, such as, O, OH, NH, NH₂, C(O), S(O)₂,C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH, S(O)₂NH₂, NHC(O)NH₂,NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂, C(O)NHS(O)₂NH orC(O)NHS(O)₂NH₂, and the like, groups comprising one or more functionalgroups, non-aromatic hydrocarbons (optionally substituted), and groupswherein one or more carbons of a non-aromatic hydrocarbon (optionallysubstituted) is replaced by a functional group. Carbon atoms of analiphatic group can be optionally oxo-substituted. An aliphatic groupmay be straight chained, branched, cyclic, or a combination thereof andpreferably contains between about 1 and about 24 carbon atoms, moretypically between about 1 and about 12 carbon atoms. In addition toaliphatic hydrocarbon groups, as used herein, aliphatic groups expresslyinclude, for example, alkoxyalkyls, polyalkoxyalkyls, such aspolyalkylene glycols, polyamines, and polyimines, for example. Aliphaticgroups may be optionally substituted. A linear aliphatic group is anon-cyclic aliphatic group. It is to be understood that when analiphatic group or a linear aliphatic group is said to “contain” or“include” or “comprise” one or more specified functional groups, thealiphatic group can be selected from one or more of the specifiedfunctional groups or a combination thereof, or a group wherein one ormore carbons of a non-aromatic hydrocarbon (optionally substituted) isreplaced by a specified functional group. In some examples, the linearaliphatic group can be represented by the formula M-V′-M′, where M andM′ are each independently absent or an alkyl, alkenyl or alkynyl, eachoptionally substituted, and V′ is a functional group. In some examples,V′ is selected from the group consisting of C(O), S(O)₂, C(O)O,C(O)N(R¹¹), OC(O)O, OC(O)N(R¹¹), S(O)₂N(R¹¹), N(R¹¹)C(O)N(R¹¹),N(R¹¹)C(O)C(O)N(R¹¹), N(R¹¹)S(O)₂N(R¹¹), C(O)N(R¹¹)S(O)₂ orC(O)N(R¹¹)S(O)₂N(R¹¹); wherein R¹¹ is as previously defined. In anotheraspect of the invention, an exemplary linear aliphatic group is analkyl, alkenyl or alkynyl, each optionally substituted, which isinterrupted or terminated by a functional group such as describedherein.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or bicyclic saturated carbocyclic ring compound by theremoval of a single hydrogen atom, and the carbon atoms may beoptionally oxo-substituted. Examples include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl,and bicyclo[2.2.2]octyl. Such alicyclic groups may be furthersubstituted.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused system, where (i) each ring system contains atleast one heteroatom independently selected from oxygen, sulfur andnitrogen, (ii) each ring system can be saturated or unsaturated, (iii)the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) thenitrogen heteroatom may optionally be quaternized, (v) any of the aboverings may be fused to an aromatic ring, and (vi) the remaining ringatoms are carbon atoms which may be optionally oxo-substituted.Representative heterocycloalkyl groups include, but are not limited to,1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl,morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl,pyridazinonyl, and tetrahydrofuryl. Such heterocyclic groups may befurther substituted. Heteroaryl or heterocyclic groups can be C-attachedor N-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphaticmoiety or the like, described herein can also be a divalent ormultivalent group when used as a linkage to connect two or more groupsor substituents, which can be at the same or different atom(s).

The term “substituted” when used with alkyl, alkenyl, alkynyl,alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, oraliphatic as described herein refers to substitution by independentreplacement of one, two, or three or more of the hydrogen atoms thereonwith substituents including, but not limited to, —F, —Cl, —Br, —I, —OH,protected hydroxy, —NO₂, —N₃, —CN, —NH₂, protected amino, oxo, thioxo,—NH—C₁-C₁₂-alkyl, —NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl,—NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl,-dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl,—O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl,—O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl,—C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl,—C(O)-aryl, —C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂,—CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl, —CONH—C₂-C₈-alkynyl,—CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl,—CONH-heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₈-alkenyl,—OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl,—OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —OCO₂—C₁-C₁₂ alkyl, —CO₂—C₂-C₈alkenyl, —CO₂—C₂-C₈ alkynyl, CO₂—C₃-C₁₂-cycloalkyl, —CO₂— aryl,CO₂-heteroaryl, CO₂-heterocyloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH— heterocycloalkyl, —NHC(O)H,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl,—NHCO₂—C₂-C₈-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₈-alkenyl,—NHC(O)NH—C₂-C₈-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₈-alkenyl,—NHC(S)NH—C₂-C₈-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₈-alkenyl,—NHC(NH)NH—C₂-C₈-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl,—NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH—C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH— heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl. It is understood that the aryls, heteroaryls, alkyls,and the like can be further substituted.

The term “halogen,” as used herein, refers to an atom selected fromfluorine, chlorine, bromine and iodine.

The term “hydrogen” includes hydrogen and deuterium. In addition, therecitation of an atom includes other isotopes of that atom so long asthe resulting compound is pharmaceutically acceptable.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl,chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl,methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl,benzyl, triphenylmethyl(trityl), methoxymethyl, methylthiomethyl,benzyloxymethyl, 2-(trimethylsilyl)ethoxymethyl, methanesulfonyl,trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “carbonyl protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a carbonyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the carbonyl protecting group as described hereinmay be selectively removed. Carbonyl protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of carbonyl protecting groups include acetals,ketals, cyclic acetals, cyclic ketals, mono- or dithioacetals, mono- ordithioketals, optionally substituted hydrazones or oximes.

The term “protected carbonyl,” as used herein, refers to a carbonylgroup protected with a carbonyl protecting group, as defined above,including dimethyl acetal, 1,3-dioxolane, 1,3-dioxane,S,S′-dimethylketal, 1,3-dithiane, 1,3-dithiolane, 1,3-oxathiolane,N,N-dimethylhydrazone, oxime, for example.

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “substituted amino,” as used herein, refers to substitution byreplacement of one or two hydrogen atoms of —NH₂ with substituentsindependently selected from the group consisting of optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted aryl, optionally substitutedcycloalkyl, optionally substituted heteroaryl, and optionallysubstituted heterocyclic; alternatively, when disubstituted, the twosubstitutents can be optionally taken together with the nitrogen atom towhich they are attached to form an optionallysubstituted heterocyclicgroup.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; hydroxy; imidazolyl; and acyloxy groups, such asacetoxy, trifluoroacetoxy and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The term “protic solvent,” as used herein, refers to a solvent thattends to provide protons, such as an alcohol, for example, methanol,ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Suchsolvents are well known to those skilled in the art, and it will beobvious to those skilled in the art that individual solvents or mixturesthereof may be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject” as used herein refers to an animal. Preferably theanimal is a mammal. More preferably the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals with undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of thepresent invention. “Prodrug”, as used herein means a compound which isconvertible in vivo by metabolic means (e.g. by hydrolysis) to acompound of the invention. Various forms of prodrugs are known in theart, for example, as discussed in Bundgaard, (ed), Design of Prodrugs,Elsevier (1985); Widder, et al. (ed), Methods in Enzymology, vol. 4,Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design andApplication of Prodrugs, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug DeliverReviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285et seq. (1988); Higuchi and Stella (eds)

Prodrugs as Novel Drug Delivery Systems, American Chemical Society(1975); and Bernard Testa & Joachim Mayer, “Hydrolysis In Drug AndProdrug Metabolism: Chemistry, Biochemistry And Enzymology,” John Wileyand Sons, Ltd. (2002).

The present invention also relates to solvates of the compounds ofFormula (I), for example hydrates.

This invention also encompasses pharmaceutical compositions containing,and methods of treating viral infections through administering,pharmaceutically acceptable prodrugs of compounds of the invention. Forexample, compounds of the invention having free amino, amido, hydroxy orcarboxylic groups can be converted into prodrugs. Prodrugs includecompounds wherein an amino acid residue, or a polypeptide chain of twoor more (e.g., two, three or four) amino acid residues is covalentlyjoined through an amide or ester bond to a free amino, hydroxy orcarboxylic acid group of compounds of the invention. The amino acidresidues include but are not limited to the 20 naturally occurring aminoacids commonly designated by three letter symbols and also includes4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,citrulline, homocysteine, homoserine, ornithine and methionine sulfone.Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, phosphate esters, dimethylaminoacetates, andphosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amine and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem., 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminun hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings and other coatings well known in thepharmaceutical formulating art. They may optionally contain opacifyingagents and can also be of a composition that they release the activeingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embeddingcompositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to VanDevanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of whichare incorporated herein by reference). A discussion of pulmonarydelivery of antibiotics is also found in U.S. Pat. No. 6,014,969,incorporated herein by reference.

Antiviral Activity

An inhibitory amount or dose of the compounds of the present inventionmay range from about 0.01 mg/Kg to about 500 mg/Kg, alternatively fromabout 0.1 to about 50 mg/Kg. Inhibitory amounts or doses will also varydepending on route of administration, as well as the possibility ofco-usage with other agents.

According to the methods of treatment of the present invention, viralinfections are treated or prevented in a patient such as a human oranother animal by administering to the subject a therapeuticallyeffective amount of a compound of the invention, in such amounts and forsuch time as is necessary to achieve the desired result. An additionalmethod of the present invention is the treatment of biological sampleswith an inhibitory amount of a compound of composition of the presentinvention in such amounts and for such time as is necessary to achievethe desired result.

The term “therapeutically effective amount” of a compound of theinvention, as used herein, means an amount of the compound which confersa therapeutic effect on the treated subject, at a reasonablebenefit/risk ratio applicable to any medical treatment. The therapeuticeffect may be objective (i.e., measurable by some test or marker) orsubjective (i.e., subject gives an indication of or feels an effect).

The term “inhibitory amount” of a compound of the present inventionmeans a sufficient amount to decrease the viral load in a biologicalsample or a subject (e.g., resulting in at least 10%, preferably atleast 50%, more preferably at least 80%, and most preferably at least90% or 95%, reduction in viral load). It is understood that when saidinhibitory amount of a compound of the present invention is administeredto a subject it will be at a reasonable benefit/risk ratio applicable toany medical treatment as determined by a physician. The term “biologicalsample(s),” as used herein, means a substance of biological originintended for administration to a subject. Examples of biological samplesinclude, but are not limited to, blood and components thereof such asplasma, platelets, subpopulations of blood cells and the like; organssuch as kidney, liver, heart, lung, and the like; sperm and ova; bonemarrow and components thereof; or stem cells. Thus, another embodimentof the present invention is a method of treating a biological sample bycontacting said biological sample with an inhibitory amount of acompound or pharmaceutical composition of the present invention.

Effective doses will also vary depending on route of administration, aswell as the possibility of co-usage with other agents. It will beunderstood, however, that the total daily usage of the compounds andcompositions of the present invention will be decided by the attendingphysician within the scope of sound medical judgment. The specifictherapeutically effective dose level for any particular patient willdepend upon a variety of factors including the disorder being treatedand the severity of the disorder; the activity of the specific compoundemployed; the specific composition employed; the age, body weight,general health, sex and diet of the patient; the time of administration,route of administration, and rate of excretion of the specific compoundemployed; the duration of the treatment; drugs used in combination orcontemporaneously with the specific compound employed; and like factorswell known in the medical arts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

When the compositions of this invention comprise a combination of acompound of the Formula (I) described herein and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 1 to 100%, andmore preferably between about 5 to 95% of the dosage normallyadministered in a monotherapy regimen. The additional agents may beadministered separately, as part of a multiple dose regimen, from thecompounds of this invention. Alternatively, those agents may be part ofa single dosage form, mixed together with the compounds of thisinvention in a single composition.

The said “additional therapeutic or prophylactic agents” include, butare not limited to, immune therapies (e.g. interferon), therapeuticvaccines, antifibrotic agents, anti-inflammatory agents such ascorticosteroids or NSAIDs, bronchodilators such as beta-2 adrenergicagonists and xanthines (e.g. theophylline), mucolytic agents,anti-muscarinics, anti-leukotrienes, inhibitors of cell adhesion (e.g.ICAM antagonists), anti-oxidants (eg N-acetylcysteine), cytokineagonists, cytokine antagonists, lung surfactants and/or antimicrobialand anti-viral agents (eg ribavirin and amantadine). The compositionsaccording to the invention may also be used in combination with genereplacement therapy.

Combination and Alternation Therapy for HCV

It has been recognized that drug-resistant variants of HCV can emergeafter prolonged treatment with an antiviral agent. Drug resistance mosttypically occurs by mutation of a gene that encodes for a protein suchas an enzyme used in viral replication, and most typically in the caseof HCV, RNA polymerase, protease, or helicase.

Recently, it has been demonstrated that the efficacy of a drug against aviral infection, such as HIV, can be prolonged, augmented, or restoredby administering the drug in combination or alternation with a second,and perhaps third, antiviral compound that induces a different mutationfrom that caused by the principal drug. Alternatively, thepharmacokinetics, biodistribution, or other parameter of the drug can bealtered by such combination or alternation therapy. In general,combination therapy is typically preferred over alternation therapybecause it induces multiple simultaneous stresses on the virus.

A compound of the present invention can also be administered incombination or alternation with antiviral agent. Examplary antiviralagents include ribavarin, interferon, interleukin or a stabilizedprodrug of any of them. More broadly described, the compound can beadministered in combination or alternation with any of the anti-HCVdrugs listed in Table 11 below.

TABLE 11 Table of anti-Hepatitis C Compounds in Current ClinicalDevelopment Drug name Drug category Pharmaceutical Company PEGASYS Longacting interferon Roche pegylated interferon alfa-2a INFERGEN Longacting interferon InterMune interferon alfacon-1 OMNIFERON Long actinginterferon Viragen natural interferon ALBUFERON Long acting interferonHuman Genome Sciences REBIF Interferon Ares-Serono interferon beta-laOmega Interferon Interferon BioMedicine Oral Interferon alpha OralInterferon Amarillo Biosciences Interferon gamma-lb Anti-fibroticInterMune IP-501 Anti-fibrotic InterMune Merimebodib VX-497 IMPDHinhibitor Vertex (inosine monophosphate dehydrogenase) AMANTADINE BroadAntiviral Agent Endo Labs (Symmetrel) Solvay IDN-6556 Apotosisregulation Idun Pharma. XTL-002 Monclonal Antibody XTL HCV/MF59 VaccineChiron CIVACIR Polyclonal Antibody NABI Therapeutic vaccine InnogeneticsVIRAMIDINE Nucleoside Analogue ICN ZADAXIN (thymosin alfa-1)Immunomodulator Sci Clone CEPLENE (histamine) Immunomodulator Maxim VX950/LY 570310 Protease inhibitor Vertex/Eli Lilly ISIS 14803 AntisenseIsis Pharmaceutical/Elan IDN-6556 Caspase inhibitor Idun PharmaceuticalsJTK 003 Polymerase Inhibitor AKROS Pharma Tarvacin Anti-PhospholipidTherapy Peregrine HCV-796 Polymerase Inhibitor ViroPharma/Wyeth CH-6Protease inhibitor Schering ANA971 Isatoribine ANADYS ANA245 IsatoribineANADYS CPG 10101 (Actilon) Immunomodulator Coley Rituximab (Rituxam)Anti-CD2O Genetech/IDEC Monoclonal Antibody NM283 (Valopicitabine)Polymerase Inhibitor Idenix Pharmaceuticals HepX ™-C Monoclonal AntibodyXTL IC41 Therapeutic Vaccine Intercell Medusa Interferon Longer actinginterferon Flamel Technology E-1 Therapeutic Vaccine InnogeneticsMultiferon Long Acting Interferon Viragen BILN 2061 Protease inhibitorBoehringer-Ingelheim TMC435350 Protease inhibitor Tibotec/MedivirTelaprevir (VX-950) Protease inhibitor Vertex Boceprevir (SCH 503034)Protease inhibitor Schering-Plough ACH-1625 Protease inhibitor AchillionABT-450 Protease inhibitor Abbott/Enanta BI-201335 Protease inhibitorBoehringer-Ingelheim PHX-1766 Protease inhibitor Phenomix VX-500Protease inhibitor Vertex MK-7009 protease inhibitor Merck R7227(ITMN-191) protease inhibitor InterMune Narlaprevir (SCH 900518)Protease inhibitor Schering/Merck Alinia (nitazoxanide) To be determinedRomark ABT-072 Polymerase Inhibitor Abbott ABT-333 Polymerase InhibitorAbbott Filibuvir (PF-00868554) Polymerase Inhibitor Pfizer VCH-916Polymerase Inhibitor Vertex R7128 (PSI6130) Polymerase InhibitorRoche/Pharmasset IDX184 Polymerase Inhibitor Idenix R1626 Polymeraseinhibitor Roche MK-3281 Polymerase inhibitor Merck PSI-7851 Polymeraseinhibitor Pharmasset ANA598 Polymerase inhibitor Anadys PharmaceuticalsBI-207127 Polymerase inhibitor Boehringer-Ingelheim GS-9190 Polymeraseinhibitor Gilead VCH-759 Polymerase Inhibitor Vertex Clemizole NS4Binhibitor Eiger Biopharmaceuticals A-832 NS5A inhibitorArrowTherapeutics BMS-790052 NS5A inhibitor Bristol-Myers-Squibb ITX5061Entry inhibitor iTherx GS-9450 Caspase inhibitor Gilead ANA773 TLRagonist Anadys CYT107 immunomodulator Cytheris SPC3649(LNA-antimiR ™-122) microRNA Santaris Pharma Debio 025 Cyclophilininhibitor Debiopharm SCY-635 Cyclophilin inhibitor Scynexis

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one of ordinary skill in theart. All publications, patents, published patent applications, and otherreferences mentioned herein are hereby incorporated by reference intheir entirety.

Abbreviations

Abbreviations which may be used in the descriptions of the scheme andthe examples that follow are: Ac for acetyl; AcOH for acetic acid; AIBNfor azobisisobutyronitrile; BINAP for2,2′-bis(diphenylphosphino)-1,1′-binaphthyl; Boc₂O fordi-tert-butyl-dicarbonate; Boc for t-butoxycarbonyl; Bpoc for1-methyl-1-(4-biphenylyl)ethyl carbonyl; BtOH for1-hydroxy-benzotriazole; Bz for benzoyl; Bn for benzyl; BocNHOH fortert-butyl N-hydroxycarbamate; t-BuOK for potassium tert-butoxide;Bu₃SnH for tributyltin hydride; BOP for(benzotriazol-1-yloxy)tris(dimethylamino)phos-phoniumHexafluorophosphate; Brine for sodium chloride solution in water; Cbzfor carbobenzyloxy; CDI for carbonyldiimidazole; CH₂Cl₂ fordichloromethane; CH₃ for methyl; CH₃CN for acetonitrile; Cs₂CO₃ forcesium carbonate; CuCl for copper (I) chloride; CuI for copper (I)iodide; dba for dibenzylidene acetone; dppb for diphenylphosphinobutane; DBU for 1,8-diazabicyclo[5.4.0]undec-7-ene; DCC forN,N′-dicyclohexylcarbodiimide; DEAD for diethylazodicarboxylate; DIADfor diisopropyl azodicarboxylate; DIPEA or (i-Pr)₂EtN forN,N-diisopropylethyl amine; Dess-Martin periodinane for1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one; DMAP for4-dimethylaminopyridine; DME for 1,2-dimethoxy-ethane; DMF forN,N-dimethylformamide; DMSO for dimethyl sulfoxide; DMT fordi(p-methoxyphenyl)phenylmethyl or dimethoxytrityl; DPPA fordiphenylphosphoryl azide; EDC forN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide; EDC HCl forN-(3-dimethylamino-propyl)-N′-ethylcarbodiimide hydrochloride; EtOAc forethyl acetate; EtOH for ethanol; Et₂O for diethyl ether; Fmoc for9-fluorenylmethoxycarbonyl; HATU forO-(7-azabenzotriazol-1-yl)-N,N,N′,N′,-tetramethyluroniumHexafluorophosphate; HCl for hydrogen chloride; HOBT for1-hydroxybenzotriazole; K₂CO₃ for potassium carbonate; n-BuLi forn-butyl lithium; i-BuLi for i-butyl lithium; t-BuLi for t-butyl lithium;PhLi for phenyl lithium; LDA for lithium diisopropylamide; LiTMP forlithium 2,2,6,6-tetramethylpiperidinate; MeOH for methanol; Mg formagnesium; MOM for methoxymethyl; Ms for mesyl or —SO₂—CH₃; Ms₂O formethanesulfonic anhydride or mesyl-anhydride; NaBH₄ for sodiumborohydride; NaBH₃CN for sodium cyanoborohydride; NaN(TMS)₂ for sodiumbis(trimethylsilyl)amide; NaCl for sodium chloride; NaH for sodiumhydride; NaHCO₃ for sodium bicarbonate or sodium hydrogen carbonate;Na₂CO₃ sodium carbonate; NaOH for sodium hydroxide; Na₂SO₄ for sodiumsulfate; NaHSO₃ for sodium bisulfite or sodium hydrogen sulfite; Na₂S₂O₃for sodium thiosulfate; NH₂NH₂ for hydrazine; NH₄HCO₃ for ammoniumbicarbonate; NH₄Cl for ammonium chloride; NMMO for N-methylmorpholineN-oxide; NaIO₄ for sodium periodate; Ni for nickel; OH for hydroxyl;OsO₄ for osmium tetroxide; Pd for palladium; Ph for phenyl; PMB forp-methoxybenzyl; POPd for dihydrogendichlorobis(di-tert-butylphosphinito-κP)palladate(II); Pd₂(dba)₃ fortris(dibenzylidene-acetone) dipalladium (0); Pd(PPh₃)₄ fortetrakis(triphenylphosphine)palladium (0); PdCl₂(PPh₃)₂ fortrans-dichlorobis(triphenyl-phosphine)palladium (II); Pt for platinum;R^(h) for rhodium; rt for room temperature; R^(u) for ruthenium; SEM for(trimethylsilyl)ethoxymethyl; TBAF for tetrabutylammonium fluoride; TBSfor tent-butyl dimethylsilyl; TEA or Et₃N for triethylamine; Teoc for2-trimethylsilyl-ethoxy-carbonyl; TFA for trifluoroacetic acid; THF fortetrahydrofuran; TMEDA for N,N,N′,N′-tetramethylethylenediamine; TPP orPPh₃ for triphenyl-phosphine; Troc for 2,2,2-trichloroethyl carbonyl; Tsfor tosyl or —SO₂—C₆H₄CH₃; Ts₂O for tolylsulfonic anhydride ortosyl-anhydride; TsOH for p-tolylsulfonic acid; TMS for trimethylsilyl;or TMSCl for trimethylsilyl chloride.

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared. Starting materials can be obtained from commercial sources orprepared by well-established literature methods known to those ofordinary skill in the art. It will be readily apparent to one ofordinary skill in the art that the compounds defined above can besynthesized by substitution of the appropriate reactants and agents inthe syntheses shown below. It will also be readily apparent to oneskilled in the art that the selective protection and deprotection steps,as well as the order of the steps themselves, can be carried out invarying order, depending on the nature of the variables to successfullycomplete the syntheses below. The variables are as defined above unlessotherwise noted below.

The compounds of the present invention may be prepared via severaldifferent synthetic routes from a variety of 5/6-membered fusedheteroaryl, 5-membered heteroaryl, and related intermediates. Aretro-synthesis of those title compounds include direct formation of asuitably linked core structure (5/6-membered fused heteroaryl or5-membered heteroaryl) followed by attachment of a suitable cappinggroup (such as —C(O)R⁶), plus some functional group manipulations inbetween and/or after. Various 5/6-membered fused heteroaryl or5-membered heteroaryl intermediates are known to those skilled in theart, for example see the encyclopedic volumns edited by A. R. Katrizky,et al, “Comprehensive Heterocyclic Chemistry” 1984; “ComprehensiveHeterocyclic Chemistry II” 1996; “Comprehensive Heterocyclic ChemistryIII” 2008.

A general synthesis and further elaboration of some 6-membered ringfused imidazole related intermediates are summarized in Scheme 1, inwhich Z is N or CH.

The synthesis starts from the construction of an optionally substitutedimidazopyridine or benzimidazole 1-2, which may be obtained bycondensation of an amino acid or its derivative 1-1.1 or 1-1.2 and a2,3-diaminopyridine or 1,2-diaminobenzene 1-1 under the conditions tothose skilled in the art. The imidazole ring closure may be realizedeither in one pot by heat, optionally in the presence of an acid and/orwith a dehydration reagent such as polyphosphoric acid; or in twosteps: 1) amide formation between diamine 1-1 and amino acid 1-1.1 or1-1.2 in the presence of a condensation reagent such as EDC HCl, DCC orthe like; or through mixed anhydride approach by reacting acid 1-1.1 or1-1.2 with a chloroformate such as methyl chloroformate, isobutylchloroformate, or the like, in the presence of a base such as TEA,DIPEA, DMAP, N-methylmorpholine, or the like, followed by treating themixed anhydride with diamine 1-1; and 2) the heterocyclic ring closurein the presence of an acid such as acetic acid, sulfuric acid or thelike or a dehydration reagent such as HATU or the like, optionally withheat.

Optionally, the NH group in the newly formed imidazopyridine orbenzimidazole ring of 1-2 may be protected with an amino protectinggroup, such as SEM (i.e. SEM-Cl, NaH), Boc, Cbz, Teoc, Troc, or thelike. The protected imidazopyridine or benzimidazole 1-2 may besubjected to lithium-halogen exchange with various (n-, s-, or t-) butyllithium and the resulting lithiate can be trapped with a nucleophile,i.e. a halide such as various allyl halide to give the allylated 1-6 asa key intermediate. Alternatively, 1-6 may be obtained from the Stillereaction conditions to those skilled in the art (see reviews: A.Anastasia, et al, Handbook of Organopalladium Chemistry for OrganicSynthesis 2002, 1, 311; F. Bellina, et al, Synthesis 2004, 2419; M. G.Organ, et al, Synthesis 2008, 2776; A. T. Lindhardt, et al, Chem.—AEuropean J. 2008, 14, 8756; E. A. B. Kantchev, et al, Angew. Chem. Int.Ed. 2007, 46, 2768; V. Farina, et al, Advances in Metal-Organic Chem.1996, 5, 1), using an allylstanne such as allyltributylstanne as theallyl donor. Analogously a key vinyl intermediates 1-3 may be preparedby Stille reaction from bromide 1-2 with tributylvinylstanne. Also,Sonogashira coupling between bromide 1-2 and propargyl alcohol ortrimethylsilyl-acetylene can generate propargyl alcohol 1-4 or alkyne1-5 after removal of TMS. Further bromination of intermediate 1-4 mayform the propargyl bromide 1-9. In addition, the bromide 1-2 may beconverted to methyl ketone 1-7 by coupling withtributyl(1-ethoxyvinyl)tin under Stille coupling conditions followed byacidic hydrolysis.

Further elaboration of the imidazopyridine or benzimidazoleintermediates starts from the vinyl intermediate 1-3, which may betransformed to aldehyde 1-8 through ozonolysis or periodate/OsO₄cleavage or to alcohol 1-12 by hydroboration-oxidation sequence. Alcohol1-12 may be converted to bromide 1-15 by the well-known brominationprocedure, which can be further functionalized to amine 1-20 throughazide substitution followed by reduction. Aldehyde 1-8 can then eitherbe reduced to alcohol 1-11, or be converted to α,β-unsaturated acid 1-10through Horner-Wadsworth-Emmons aldehyde homologation reaction followedby saponification. Alcohol 1-11 may be similarly converted to thecorresponding amine intermediate 1-14 and bromide intermediate 1-13 asdescribed previously. Bromide 1-13 can be homologated to alkyneintermediate 1-19 with a metal acetylide. In addition, bromide 1-13 maybe also transformed to thiol 1-16 through nucleophilic substitution,which can be further oxidized to sulfonic acid 1-17. Sulfonamide 1-18may then be derived from 1-17 through the sulfonyl chloride activationprocess.

It should be noted that optionally the NH group of all theimidazopyridine or benzimidazole related intermediates listed above maybe protected with an amino protecting group, such as SEM (i.e. SEM-Cl,NaH), Boc, Cbz, Teoc, Troc, or the like.

A typical synthesis of an imidazole related intermediate is analogous tothat of the imidazopyridine or benzimidazole intermediates. As shown inScheme 2, bromo-imidazole 2-4 can be synthesized in a three-stepsequence: 1) condensation between amino acid derived aldehyde 2-1.1 or2-1.2 and glyoxal 2-1.3 in the presence of methanolic ammonia togenerate imidazole 2-2; 2) bromination of 2-2 with excess amount ofbromination reagent such as 2,4,4,6-tetrabromo-2,5-cyclohexadienone,NBS, etc. to afford dibromide 2-3; and 3) selective reduction of thedibromide 2-3 by heating in aq. Na₂SO₃ or aq. NaHSO₃. 2-4 then may beserved as a universal intermediate further elaborable to many otherimidazole derivatives using the chemistry discussed in Scheme 1, some ofwhich are listed in the table below.

TABLE 12

Optionally, the NH group of imidazole related intermediates listed abovemay be protected with an amino protecting group (shown in Scheme 2 asPG, theoretically the reaction will generate two regio-isomers, but onlyone is drawn for clarity), such as SEM (i.e. SEM-Cl, NaH), Boc, Cbz,Teoc, Troc, or the like. The protected imidazole 2-5 may be deprotonatedwith a strong base such as LDA, BuLi, etc to generate a carbon anion,which may either undergo a nucleophilic substitution with an activatedhalide such as 2-5.2 to afford aryl or heteroaryl substituted imidazole2-6 or couple with an aryl or heteroaryl halide 2-5.1 in the presenceappropriate transition metal salt to generate bicyclic heteroaryl 2-7.Similarly, the protected bromo imidazole 2-8 may be subjected tolithium-halogen exchange with various (n-, s-, or t-) butyl lithium, theresulting lithiate may undergo similar reactions to afford 2-6 and 2-7.Also, when 2-8 is treated with metallated aryl or heteroaryl 2-8.1, inwhich M at each occurrence is independently a boron, tin, silicon, zinc,zirconium, or copper species, under Suzuki, Stille or related couplingconditions known to those skilled in the art (see reviews: A. Suzuki,Pure Applied Chem., 1991, 63, 419; A. Suzuki, Handbook ofOrganopalladium Chemistry for Organic Synthesis, 2002, 1, 249; A.Anastasia, et al, Handbook of Organopalladium Chemistry for OrganicSynthesis 2002, 1, 311; F. Bellina, et al, Synthesis, 2004, 2419; M. G.Organ, et al, Synthesis, 2008, 2776; A. T. Lindhardt, et al, Chem.—AEuropean J., 2008, 14, 8756; E. A. B. Kantchev, et al, Angew. Chem. Int.Ed., 2007, 46, 2768; V. Farina, et al, Advances in Metal-Organic Chem.,1996, 5, 1), to provide coupling product 2-7. In addition to thesedirect coupling strategy, aryl or heteroaryl bromide 2-5.1 may beconverted to methyl ketone 2-9 under Stille coupling conditions withtributyl(1-ethoxyvinyl)tin 2-9.1.2-9 may be brominated under conditionsto those skilled in the art to afford bromide 2-10, which may be eitherconverted to the corresponding amine 2-11, or coupled with protectedamino acid 1-1.1 or 1-1.2 in the presence of a base such as Et₃N andDIPEA to afford keto-ester 2-12. Similarly, amine 2-11 may be convertedto the corresponding keto-amide 2-13 via condensation with appropriateamino acid under standard amide formation conditions. 2-12 and 2-13 maybe transformed to key intermediate 2-14 via heating with (NH₄)OAc underthermal or microwave conditions.

Similarly when Q is a fused bicyclic group as previously defined,compounds of the present invention may be synthesized from intermediates3-2 and 3-3 (as shown in Scheme 3), which may be synthesized usingsimilar chemistry described in Schemes 1 and 2 from the bicycliccarboxylic acid 3-1. Bicyclic carboxylic acids or derivatives withvarious structural features are either commercially available or knownin the literature, and have been extensively reviewed by S. Hanessian,et al (Tetrahedron, 1997, 53, 12789) and A. Trabocchi, et al (AminoAcids, 2008, 34, 1) which are incorporated herein by reference.

Other requisite buiding blocks for the syntheses of the compounds of thepresent invention are various amide, carbamate or urea-relatedintermediates, which can be prepared from a suitable, commerciallyavailable amine through standard amide, carbamate or urea formation bydirect condensation of an carboxylic acid with a dehydration and/orcondensation reagent, such as HATU, DCC, BtOH, EDC or the like; or achloroformate; or an isocyanate; in the presence of a suitable base suchas pyridine, TEA, DIPEA, DMAP, NaHCO₃, K₂CO₃ or the like. For example,as shown in Scheme 3a, the acid 3-1a can be condensed with4-bromoaniline 3-2a in the presence of HATU and DIPEA in CH₂Cl₂ toafford amide 3-3a.

Other examples of some of the amide-related intermediates that may beused to construct the title compounds of the present invention arecompiled in the table below.

TABLE 13

With a variety of suitably substituted imidazopyridines, benzimidazolesand imidazoles such as those listed in Schemes 1-3 and the amide-relatedderivatives such as that in Scheme 3a, and the tables above in hand, thecompounds of the present invention may be prepared through variouscoupling strategy or a combination of strategies to connect twofragments, optionally with a suitable cyclic or acyclic linker orformation of a cyclic or acyclic linker. The said strategy includes, butnot limited to, Stille coupling, Suzuki coupling, Sonogashira coupling,Heck coupling, Buchwald amidation, Buchwald amination, Williametherification, Buchwald etherification, alkylation, pericyclic reactionwith different variations, or the like.

An example of the strategies that may be used to prepare the compoundsof the present invention is shown in Scheme 4. Both bromides 4-1 and 4-2can be prepared using similar procedures described in Schemes 1-3 and3a. Bromide 4-2 can be converted to the corresponding metallated aryl4-3 (boronate or stanne) under Suzuki or Stille conditions withbis(pinacolato)diboron, hexamethylditin or hexabutylditin in thepresence of Pd-catalyst. The latter may be further coupled withimidazopyridine bromide 4-1 under similar conditions to generate a corestructure 4-4.

Compound 4-4 then may be served as a common intermediate for furtherderivatizations to 4-5 in two steps: 1) mono-deprotection of the linearor cyclic amine moiety may be accomplished, for example, treatment tohydrogenolytic conditions under Pd catalyst to remove the Cbz protectiongroup; and 2) the released amine functionality may be acylated with ancarboxylic acid under standard acylation conditions, for example acoupling reagent such as HATU in combination with an organic base suchas DIPEA can be used in this regard; alternatively, the released aminemay be reacted with an isocyanate, carbamoyl chloride or chloroformateto provide an urea or carbamate. Various carboxylic acids includingamino acids in racemic or optical form are commercially available,and/or can be synthesized in racemic or optical form, see referencescited in reviews by D. Seebach, et al, Synthesis, 2009, 1; C. Cativielaand M. D. Diaz-de-Villegas, Tetrahedron: Asymmetry, 2007, 18, 569; 2000,11, 645; and 1998, 9, 3517; and experimental examples compiled in patentapplication WO 08/021927 A2 by C. Bachand, et al, from Bristol-MyersSquibb, Co., which is incorporated herein by reference. 4-5 may befurther deprotected under hydrolytic conditions in the presence of anacid such as TFA or hydrogen chloride to remove the Boc protection groupand the released amine functionality can be further derivatized to thetitle compounds I-1, with a carboxylic acid using the conditionsdescribed above.

Alternatively, as shown in Scheme 4a, the compounds of the presentinvention (for example I-1) may also be derived frombromoimidazopyridine or bromobenzimidazole 4-1a and amide 4-2a using theprocedures described previously. The intermediates 4-1a and 4-2a havethe desired acyl groups already installed using similar sequences shownin Scheme 4.

The compounds of the present invention containing five-memberedheteroaryl other than imidazole may be prepared using similar proceduresdescribed above in Schemes 1-4 and 4a. For example, some intermediatescontaining a desired, suitably substituted five-membered heteroaryl havebeen published in US 2008/0311075A1 by C. Bachand, et al fromBristol-Myers Squibb, Co., which is incorporated by reference. Thesesintermediates are compiled in the following table.

TABLE 14

The synthesis of the compounds of the present invention involves5/6-membered fused heteroaryl intermediates other than imidazopyridinesor benzimidazoles, various 5/6-membered fused heteroaryl are known inthe literature. The synthesis of other 5/6-membered fused heteroarylintermediates depends on the chemical features of each structure. Forexample, a typical synthesis of indole intermediate is illustrated inScheme 5. The commercially available bromoiodoaniline 5-1 may be coupledto the commercially available acetylene 5-1.1 under the Sonogashiraconditions to give phenylacetylene 5-2. The latter may be cyclized toindole 5-3 under heat or microwave condition in the presence of a coppercatalyst.

It will be appreciated that, with appropriate manipulation andprotection of any chemical functionality, synthesis of compounds ofFormula (I) is accomplished by methods analogous to those above and tothose described in the Experimental section. Suitable protecting groupscan be found, but are not restricted to, those found in T W Greene and PG M Wuts “Protective Groups in Organic Synthesis”, 3rd Ed (1999), JWiley and Sons.

In certain aspects, the invention encompasses a process of making acompound of the invention comprising:

-   -   i) Preparing a compound of Formula (II):

-   -   via a transition-metal catalyzed cross-coupling reaction;    -   wherein:    -   G, U, R¹, R^(1a), R^(1b), R^(1c), R^(7a), R^(7b), R⁹, and R^(9a)        are as defined above;    -   Ring A¹ is absent, or optionally substituted aryl or optionally        substituted heteroaryl;    -   Ring B¹ is optionally substituted aryl or optionally substituted        heteroaryl;    -   L¹ is absent, or optionally substituted C₂-C₄ alkenyl or C₂-C₄        alkynyl; and    -   Z^(a) or Z^(b) are each independently an amino protecting group        or —C(O)—R⁶; wherein R⁶ is as defined in claim 1;    -   ii) When Z^(a) or Z^(b) is an amino protecting group, fully or        selectively deprotecting a compound of Formula (II) to give the        corresponding amine of Formula (III):

-   -   wherein Z^(c) is hydrogen, an amino protecting group or        —C(O)—R⁶;    -   iii) Capping the released amino group of a compound of        Formula (III) with LG-C(O)—R⁶,        -   wherein LG is a leaving group; to give the compound of            Formula (IV):

-   -   wherein Z^(d) is an amino protecting group or —C(O)—R⁶; and    -   iv) Repeated reaction sequence of deprotecting and capping (step        ii-iii) to give the compound of Formula (V):

All references cited herein, whether in print, electronic, computerreadable storage media or other form, are expressly incorporated byreference in their entirety, including but not limited to, abstracts,articles, journals, publications, texts, treatises, internet web sites,databases, patents, and patent publications.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

The compounds of examples 1-347 may be prepared using procedures similarto those described in example 348 (described below), and/or as describedin the Synthetic Method.

Examples 1-219

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Examples 220-229

Entry R R′ R″ U Entry R R′ R″ U 220 Me H H CH₂ 221 H H H CF₂ 222 Me H HS 223 H H H

224 Me H H O 225 H H H

226 H Ph H CH₂ 227 H H H

228 H H Ph CH₂ 229 H H H

Examples 230-233

Entry E Entry E Entry E Entry E 230 absent 231 CH₂ 232 CH₂O 233 OCH₂CH₂

Examples 234-259

Entry R 234

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Examples 260-268

En- En- try Q^(q) try Q^(q) 260

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Examples 269-278

Entry R R′ R″ R′′′ 269 F H H H 270 F F H H 271 Me H H H 272 Me Me H H273 H H Me Me 274 H H Et Et 275 CF₃ H H H 276 CF₃ H CF₃ H 277 Cl H H H278 Cl H Cl H

Examples 279-296

Entry R R′ R″ 279 Me H H 280 H CO₂H H 281 H F H 282 H H CO₂H 283 H H F284 H CO₂Me H 285 H Cl H 286 H H CO₂Me 287 H H Cl 288 H CONH₂ H 289 H MeH 290 H H CONH₂ 291 H H Me 292 H OMe H 293 H CF₃ H 294 H H OMe 295 H HCF₃ 296 CO₂Me H H

Examples 297-314

Entry A^(a) 297

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Examples 315-335

Entry A^(a) 315

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Examples 336-347

Entry G^(g) 336

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Example 348

Step 348a. Into a mixture of(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (486 mg, 2.26mmol) and 5-iodopyrimidin-2-amine (500 mg, 2.26 mmol) in acetonitrile(10 mL) was added diisopropylethylamine (0.45 mL, 2.48 mmol) and HATU(859 mg, 2.26 mmol). The resulting mixture was stirred at 150° C. for1.5 hours (heating with microwave) before being partitioned betweenwater and EtOAc. The organic phase was separated, dried (Na₂SO₄) andconcentrated to afford a brown slurry, which was purified by flashcolumn chromatography (silica, EtOAc-hexanes) to give the desiredproduct as a light yellow solid (146 mg, 15%). ESIMS m/z=440.93 [M+Na]⁺.

Step 348b. 6-bromo-N-methoxy-N-methyl-2-naphthamide (prepared accordingto J. Med. Chem., 2006, 49, 4721-4736, 3.57 g, 12.1 mmol) in THF (60 mL)was treated with methyl magnesium bromide (3M in Et₂O, 8.09 mL, 24.3mmol) slowly at 0° C. for 1 hour. The solution was warmed up to rt for 2hours before being quenched with aqueous NH₄Cl. The volatiles wereremoved and the residue was partitioned (EtOAc-water). The organics werewashed with brine, dried (Na₂SO₄), filtered and evaporated to give thecrude desired compound as a white solid (2.89 g, 96%).

Step 348c. The compound from step 348b (2.89 g, 11.6 mmol) in aceticacid (60 mL) was treated with bromine (0.59 mL, 11.6 mmol) dropwise for1 hour. The volatiles were evaporated and the residue was partitioned(EtOAc—saturated aqueous NaHCO₃). The organics were washed with brine,dried (Na₂SO₄), filtered and evaporated to give the crude desiredcompound as a light yellow solid (3.898 g).

Step 348d. A mixture of the compound from step 348c (at most 11.6 mmol)and N-Boc-L-proline (3.75 g, 17.4 mmol) in CH₃CN (60 mL) was added DIPEA(2.89 mL, 23.2 mmol) slowly. The mixture was stirred at rt until thedisappearence of the starting material. The volatiles were evaporatedand the residue was partitioned (EtOAc-water). The organics were washedwith brine, dried (Na₂SO₄), filtered and evaporated to give the crudedesired compound as a yellow-white foam (4.762 g). ESIMS m/z=462.03,464.02 [M+H]⁺.

Step 348e. A mixture of the compound from step 348d (0.200 g, 0.452mmol), bis(pinacolato)diboron (0.144 g, 0.565 mmol), PdCl₂(dppf)₂ (36.9mg, 0.0452 mmol) and potassium acetate (88.7 mg, 0.904 mmol) in DMSO (5mL) was degassed and heated at 80° C. under N₂ for 17 hours. Thereaction mixture was allowed to cool down and partitioned (EtOAc-water).The organic layer was washed with brine, dried (Na₂SO₄), filtered andevaporated. The residue was purified by chromatography (silica,hexanes-ethyl acetate) to give the desired compound as a yellow solid(0.188 g, 85%). ESIMS m/z=490.12 [M+H]⁺.

Step 348f. Into a mixture of compound from step 348a (146 mg, 0.35 mmol)and the compound from step 348e (171 mg, 0.35 mmol) in1,2-dimethoxyethane (5 mL) and water (2.5 mL) was addedtetrakis(triphenylphosphine)palladium(0) (40 mg, 0.035 mmol) and NaHCO₃(371 mg, 4.42 mmol). The resulting mixture was stirred at 65° C. undernitrogen atmosphere for 1.5 hours before being partitioned between waterand EtOAc. The organic phase was separated, dried (Na₂SO₄) andconcentrated to afford a brown slurry, which was purified by flashcolumn chromatography (silica, ethyl acetate-hexanes) to give thedesired product as a light yellow solid (38 mg, 17%). ESIMS m/z=654.13[M+H]⁺.

Step 348g. A solution of the compound from step 348f (38 mg, 0.058 mmol)in dichloromethane (1 mL) and methanol (0.1 mL) was treated with HCl in1,4-dioxane (4 M, 2 mL) at room temperature for 2 hours. The volatileswere evaporated off to give the crude desired compound as a yellow solidwhich was used directly in the next step.

Step 348h. The mixture of compounds from step 348g (0.048 mml at most)and (S)-(methoxycarbonyl)amino-3-methyl-butyric acid (prepared accordingto WO 2008/021927, 20 mg, 0.11 mmol) in MeCN (2 mL) was addeddiisopropylethylamine (0.11 mL) and HATU (7.7 mg). The resultingsolution was stirred at room temperature for 35 minutes before beingpartitioned between EtOAc and aqueous NaOH (0.5M). The organic phase wasseparated, dried (Na₂SO₄) and concentrated to afford a brown slurry,which was purified by HPLC(C18, methanol-water) to give the titlecompound (2.0 mg). ESIMS m/z=768.35 [M+H]⁺.

Biological Activity

1. HCV Replicon Cell Lines

HCV replicon cell lines (kindly provided by R. Bartenschlager) isolatedfrom colonies as described by Lohman et. al. (Lohman et al. (1999)Science 285: 110-113, expressly incorporated by reference in itsentirety) and used for all experiments. The HCV replicon has the nucleicacid sequence set forth in EMBL Accession No.: AJ242651, the codingsequence of which is from nucleotides 1801 to 8406.

The coding sequence of the published HCV replicon was synthesized andsubsequently assembled in a modified plasmid pBR322 (Promega, Madison,Wis.) using standard molecular biology techniques. One replicon cellline (“SGR 11-7”) stably expresses HCV replicon RNA which consists of(i) the HCV 5′UTR fused to the first 12 amino acids of the capsidprotein, (ii) the neomycin phosphotransferase gene (neo), (iii) the IRESfrom encephalomyocarditis virus (EMCV), and (iv) HCV NS2 to NS5B genesand the HCV 3′UTR. Another replicon cell line (“Huh-luc/neo-ET”)described by Vrolijk et. al. (Vrolijk et. al. (2003) Journal ofVirological Methods 110:201-209, expressly incorporated by reference inits entirety) stably expresses HCV replicon RNA which consists of (i)the HCV 5′UTR fused to the first 12 amino acids of the capsid protein,(ii) the firefly luciferase reporter gene, (iii) the ubiquitin gene,(iv) the neomycin phosphotransferase gene (neo), (v) the IRES fromencephalomyocarditis virus (EMCV), and (vi) HCV NS3 to NS5B genes thatharbor cell culture adaptive mutations (E1202G, T1280I, K1846T) and theHCV 3′UTR.

These cell lines were maintained at 37° C., 5% CO₂, 100% relativehumidity in DMEM (Cat# 11965-084, Invitrogen), with 10% fetal calf serum(“FCS”, Invitrogen), 1% non-essential amino acids (Invitrogen), 1% ofGlutamax (Invitrogen), 1% of 100× penicillin/streptomycin (Cat#15140-122, Invitrogen) and Geneticin (Cat# 10131-027, Invitrogen) at0.75 mg/ml or 0.5 mg/ml for 11-7 and Huh-luc/neo-ET cells, respectively.

2. HCV Replicon Assay—qRT-PCR

EC₅₀ values of single agent compounds and combinations were determinedby HCV RNA detection using quantitative RT-PCR, according to themanufacturer's instructions, with a TAQMAN® One-Step RT-PCR Master MixReagents Kit (Cat# AB 4309169, Applied Biosystems) on an ABI Model 7500thermocycler. The TaqMan primers used for detecting and quantifying HCVRNA were obtained from Integrated DNA Technologies. HCV RNA wasnormalized to GAPDH RNA levels in drug-treated cells, which was detectedand quantified using the Human GAPDH Endogenous Control Mix (AppliedBiosystems, AB 4310884E). Total cellular RNA was purified from 96-wellplates using the RNAqueous 96 kit (Ambion, Cat# AM1812). Chemical agentcytotoxicity was evaluated using an MTS assay according to themanufacturer's directions (Promega).

3. HCV Replicon Assay—Luciferase

Since clinical drug resistance often develops in viral infectionsfollowing single agent therapies, there is a need to assess theadditive, antagonistic, or synergistic properties of combinationtherapies. We use the HCV replicon system to assess the potential use ofthe compound of the present invention or in combination therapies withInterferon alpha, cyclosporine analogs and inhibitors targeting otherHCV proteins. The acute effects of a single or combinations of drugs arestudied in the “Huh-luc/neo-ET” replicon with each chemical agenttitrated in an X or Y direction in a 6 point two-fold dilution curvecentered around the EC50 of each drug. Briefly, replicon cells areseeded at 7,000 cells per well in 90 ul DMEM (without phenol red,Invitrogen Cat.# 31053-036) per well with 10% FCS, 1% non-essentialamino acids, 1% of Glutamax and 1% of 100× penicillin/streptomycin andincubated overnight at 37° C., 5% CO₂, 100% relative humidity. 16-20hafter seeding cells, test compounds previously solubilized and titratedin dimethyl sulfoxide (“DMSO”) from each X plate and Y plate are diluted1:100 in DMEM (without phenol red, Invitrogen Cat.# 31053-036) with 10%FCS, 1% non-essential amino acids, 1% of Glutamax and 1% of 100×penicillin/streptomycin and added directly to the 96-well platecontaining cells and growth medium at a 1:10 dilution for a finaldilution of compound and DMSO of 1:1000 (0.2% DMSO final concentration).Drug treated cells are incubated at 37° C., 5% CO₂, 100% relativehumidity for 72 hours before performing a luciferase assay using 100 ulper well BriteLite Plus (Perkin Elmer) according to the manufacturer'sinstructions. Data analysis utilizes the method published by Prichardand Shipman (Antiviral Research, 1990. 14:181-205). Using this method,the combination data are analyzed for antagonistic, additive, orsynergistic combination effects across the entire combination surfacecreated by the diluted compounds in combination.

The compounds of the present invention may inhibit HCV by mechanisms inaddition to or other than NS5A inhibition. In one embodiment, thecompounds of the present invention inhibit HCV replicon and in anotherembodiment the compounds of the present invention inhibit NS5A.

The compounds of the present invention can be effective against the HCV1b genotype. It should also be understood that the compounds of thepresent invention can inhibit multiple genotypes of HCV. In oneembodiment compound of the present invention are active against the 1a,1b, 2a, 2b, 3a, 4a, and 5a genotypes. Table 15 shows the EC₅₀ value of arepresentative compound of the present invention against the HCV 1bgenotype from the above described qRT-PCR or luciferase assay. EC₅₀ranges against HCV 1b are as follows: A >10 nM; B1-10 nM; C<1 nM.

TABLE 15 Genotype-1b replicon EC₅₀ Example Range 348 B

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed is:
 1. A compound selected from compounds compiled inthe following table or a pharmaceutically acceptable salt thereof:

Entry

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2. A pharmaceutical composition comprising a compound or a combinationof compounds according to claim 1 or a pharmaceutically acceptable saltthereof, in combination with a pharmaceutically acceptable carrier orexcipient.
 3. A compound selected from compounds compiled in thefollowing tables or a pharmaceutically acceptable salt thereof:

Entry R R′ R″ U 220 Me H H CH₂ 221 H H H CF₂ 223 H H H

225 H H H

226 H Ph H CH₂ 227 H H H

228 H H Ph CH₂ 229 H H H


4. A compound selected from compounds compiled in the following table ora pharmaceutically acceptable salt thereof:

Entry Q^(q) 260

261

262


5. A compound selected from compounds compiled in the following table ora pharmaceutically acceptable salt thereof:

Entry R R′ R″ R′′′ 269 F H H H 270 F F H H 271 Me H H H 272 Me Me H H273 H H Me Me 274 H H Et Et 275 CF₃ H H H 276 CF₃ H CF₃ H 277 Cl H H H278 Cl H Cl H.


6. A compound selected from compounds compiled in the following table ora pharmaceutically acceptable salt thereof:

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